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Regulation of gene expression encoding the digestive α-amylase in the larvae of Colorado potato beetle, Leptinotarsa decemlineata (Say) in response to plant protein extracts
Gene ( IF 3.5 ) Pub Date : 2020-09-21 , DOI: 10.1016/j.gene.2020.145159
Shabnam Ashouri , Reza Farshbaf Pourabad

Considering the relevance of insect α-amylases and natural α-amylase inhibitors present in plants to protect against insect damage, we investigated the effect of white bean and rapeseed protein extracts on digestive α-amylase gene expression of the Colorado potato beetle, Leptinotarsa decemlineata (Say). For this purpose, in vitro and in vivo trials were performed to determine the inhibitory activity of seed proteins on the third and fourth instar larvae. In both trials, the significant inhibitory effect of each extracts on the third and fourth instar larval α-amylase activity and considerable mortality in treatments were observed compared to control trials. In the RT-qPCR, expression ratio demonstrated that the α-amylase gene of two different larval stages grown on both proteins treated leaves had significantly differentiated expression and was up-regulated in third instar larvae and down-regulated in fourth instar larvae compared to control. Results suggest that the hyper-production of α-amylase in third instar larvae is elicited to compensate for the enzyme activity inhibition at an earlier stage and also down-regulation suggests the existence of a negative feedback of plant proteins on the last instar larvae via impaired food intake and digestive α-amylase activity in Colorado potato beetle. Therefore, disruption of the insect’s digestive physiology by plant defensive proteins can be considered in the development of innovative controlling methods of this crucial potato pest.



中文翻译:

响应植物蛋白提取物,调节科罗拉多马铃薯甲虫Leptinotarsa decemlineata(Say)幼虫中编码消化α-淀粉酶的基因表达

考虑到植物中存在的昆虫α-淀粉酶和天然α-淀粉酶抑制剂具有相关性以防止昆虫受到伤害,我们研究了白豆和菜籽蛋白提取物对科罗拉多马铃薯甲虫Leptinotarsa decemlineata的消化α-淀粉酶基因表达的影响(说)。为了这个目的,进行了体外和体内试验以确定种子蛋白对第三和第四龄幼虫的抑制活性。在两个试验中,与对照试验相比,观察到每种提取物对第三和第四龄幼虫α-淀粉酶活性具有明显的抑制作用,并且在治疗中具有相当大的死亡率。在RT-qPCR中,表达率表明,与对照相比,在两种蛋白质处理的叶片上生长的两个不同幼虫阶段的α-淀粉酶基因表达差异显着,在三龄幼虫中上调,在四龄幼虫中下调。 。结果表明,第三龄幼虫幼虫中α-淀粉酶的过度产生可以补偿早期的酶活性抑制,并且下调表明,受损的末龄幼虫中存在植物蛋白的负反馈食物摄入量和科罗拉多马铃薯甲虫的消化α-淀粉酶活性。因此,在开发这种重要马铃薯害虫的创新防治方法时,可以考虑用植物防御蛋白破坏昆虫的消化生理。

更新日期:2020-09-25
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